Distinct effects of Abelson kinase mutations on myocytes and neurons in dissociated Drosophila embryonic cultures: mimicking of high temperature.
Distinct effects of Abelson kinase mutations on myocytes and neurons in dissociated Drosophila embryonic cultures: mimicking of high temperature.
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DOI：
10.1371/journal.pone.0086438
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Wu CF
中科院分区：
文献类型：
--
作者：
Liu L;Wu CF
Abelson酪氨酸激酶（Abl）在体内调节轴突引导、肌肉发育和细胞-细胞相互作用。果蝇原代培养系统为探索Abl介导的细胞机制提供了多种实验手段。在这里，我们证明了单胚胎培养在神经元和肌细胞的细胞分化和发育进展以及神经-肌肉接触方面表现出阶段依赖的特征。特别是，肌肉的发育在很大程度上取决于分离胚胎的阶段。在野生型（WT）培养中，在12期之前的胚胎中，肌肉细胞仍然在细胞簇中，很少被检测到。有趣的是，在Abl突变培养物中发现了大量的肌细胞，即使在与较年轻的10期胚胎分离的培养物中，也表现出增强的肌细胞运动和融合，以及神经元-肌肉接触。值得注意的是，Abl肌细胞经常显示扩张良好的板足。相反，Abl神经元的特征是较少的大薄纱样板足，而是有更多的丝状足和沿神经突的较暗的淋巴结。这些不同的表型在不同等位基因（Abl1和Abl4）的同型和杂合子培养（Abl/Abl vs. Abl/+）中同样明显，表明显性突变效应。引人注目的是，在10期胚胎的WT培养物中，高温（HT）培养促进了肌肉的迁移和融合，部分模仿了Abl培养物典型的高级肌肉发育。然而，与Abl神经元的特征形态不同，高温可以促进神经元的生长，增加板足的数量。有趣的是，HT孵育也促进Abl板足扩张，对神经细胞的影响远大于肌肉细胞。我们的研究结果表明，Abl是肌细胞和神经元发育的重要调节因子，高温培养部分模仿Abl培养的更快肌肉发育。尽管Abl突变引起了广泛的改变，但在标记胚胎的WT和Abl混合培养中，我们观察到WT和Abl细胞之间的肌细胞融合事件和神经肌肉接触的形成。
Abelson tyrosine kinase (Abl) is known to regulate axon guidance, muscle development, and cell-cell interaction in vivo. The Drosophila primary culture system offers advantages in exploring the cellular mechanisms mediated by Abl with utilizing various experimental manipulations. Here we demonstrate that single-embryo cultures exhibit stage-dependent characteristics of cellular differentiation and developmental progression in neurons and myocytes, as well as nerve-muscle contacts. In particular, muscle development critically depends on the stage of dissociated embryos. In wild-type (WT) cultures derived from embryos before stage 12, muscle cells remained within cell clusters and were rarely detected. Interestingly, abundant myocytes were spotted in Abl mutant cultures, exhibiting enhanced myocyte movement and fusion, as well as neuron-muscle contacts even in cultures dissociated from younger, stage 10 embryos. Notably, Abl myocytes frequently displayed well-expanded lamellipodia. Conversely, Abl neurons were characterized with fewer large veil-like lamellipodia, but instead had increased numbers of filopodia and darker nodes along neurites. These distinct phenotypes were equally evident in both homo- and hetero-zygous cultures (Abl/Abl vs. Abl/+) of different alleles (Abl1 and Abl4) indicating dominant mutational effects. Strikingly, in WT cultures derived from stage 10 embryos, high temperature (HT) incubation promoted muscle migration and fusion, partially mimicking the advanced muscle development typical of Abl cultures. However, HT enhanced neuronal growth with increased numbers of enlarged lamellipodia, distinct from the characteristic Abl neuronal morphology. Intriguingly, HT incubation also promoted Abl lamellipodia expansion, with a much greater effect on nerve cells than muscle. Our results suggest that Abl is an essential regulator for myocyte and neuron development and that high-temperature incubation partially mimics the faster muscle development typical of Abl cultures. Despite the extensive alterations by Abl mutations, we observed myocyte fusion events and nerve-muscle contact formation between WT and Abl cells in mixed WT and Abl cultures derived from labeled embryos.
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影响因子：
14.8
作者：
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通讯作者：
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影响因子：
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